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題名 | 黑殭菌MA126菌株DNA探針製備及田間偵測應用=The Preparation of DNA Probe of Metarhizium Anisopliae MA126 Isolate and Its Application in the Field |
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作者 | 劉顯達; 林秀芬; | 書刊名 | 植物保護學會會刊 |
卷期 | 49:4 2007.12[民96.12] |
頁次 | 頁347-354 |
分類號 | 433.85 |
關鍵詞 | 蟲生真菌; 黑殭菌; DNA探針; Entomopathogenic fungi; Metarhizium anisopliae; DNA probe; |
語文 | 中文(Chinese) |
中文摘要 | 我國地處亞熱帶,蟲害問題特別嚴重,在作物生產上,長久以來過分依賴與不當使用化學殺蟲劑,造成環境污染、殘毒、抗藥性及危害人體等問題,加上環保意識高漲及全球性對於地球生態永續性的重視,研究發展生物性農藥,已為時勢之所趨。真菌殺蟲劑具有對環境生態影響小,且蟲害不易產生抗藥性等優點(6),其中以黑殭菌(Metarhizium anisopliae)可寄生多達200餘種的昆蟲(25),具有開發為作物害蟲防治用生物製劑之潛力(2,17)。黑殭菌(Metarhizium)屬包含三個種,分別為小孢子型黑殭菌(M. anisopliae)和大孢子型黑殭菌(M. flavoviridae及M. album)。其中M. anisopliae比大孢子型的黑殭菌M. flavoviridae有較廣之地理分布及寄主範圍,且從基因遺傳角度來看,M. anisopliae比M. flavoviridae具有較大的變異性(3);小孢子型M. anisopliae依其分生孢子的大小,又可區分成M. anisopliae var. anisopliae小孢變種和M. anisopliae var. majus大孢變種兩類(13)。M. anisopliae var. anisopliae比M. anisopliae var. majus有較高的致病性,而且個體間也產生許多菌落顏色及孢子型態的差異(19)。因此惟準確鑑定黑殭菌特定菌株,才能為其產業利用提供智財保護(23)。 |
英文摘要 | Random amplification of polymorphic DNA polymerase chain reaction (RAPD-PCR) based technology was used to assess the genomic variability among 13 isolates of entomopathogenic fungi Metarhizium anisopliae. The high level of variability in PCR-amplified fragments within M. anisopliae MA126 was similar to the total variability between M. anisopliae MAUZ1, M. anisopliae KCAL, and M. anisopliae 35520. Using sequence information from a region of the M. anisopliae specific probe OPI-18-1.0kb, primers OPI-18-1.0 kb-LP (5'-ATATCTTCTAATAA GAGCCGTTTC) and OPI-18-1.0 kb-RP (5'-CCTGGTTCGACTGGCAATGC) were generated. This primer pair amplified similar 1.0 kb size products from four isolates of M. anisopliae, namely, M. anisopliae MA126, M. anisopliae MAUZ1, M. anisopliae KCAL, and M. anisopliae 35520. Hybridization with the probe indicated the presence of DNA homology among the M. anisopliae MA126, M. anisopliae MAUZ1, M. anisopliae KCAL, and M. anisopliae 35520. The OPI-18-1.0kb probe provides a useful and rapid tool for identifying species and strains of entomopathogenic fungus M. anisopliae in the field. |
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